An automated capillary electrophoresis system for high-speed separation of DNA fragments based on a short capillary

• Published in: Electrophoresis 2010-10, Vol.31 (19), p.3184-3191
• Main Authors: Cheng, Yong-Qiang, Yao, Bo, Zhang, Hui-Dan, Fang, Jin, Fang, Qun
• Format: Article
• Language: English
• Subjects: Automation - instrumentation; Bacteriophages - genetics; DNA - chemistry; DNA - isolation & purification; DNA fragments separation; Electrophoresis, Capillary - instrumentation; Electrophoresis, Capillary - methods; Equipment Design; High-speed CE; Molecular Weight; Polymerase Chain Reaction; Reference Standards; Reproducibility of Results; Sensitivity and Specificity; Short capillary; Time Factors
• ISSN: 0173-0835; 1522-2683; 1522-2683
• DOI: 10.1002/elps.201000362

A high‐speed DNA fragment separation system was developed based on a short capillary and a slotted‐vial array automated sample introduction system. The injection process of DNA sample in a short capillary was investigated systematically with three injection techniques including constant‐field‐strength, low‐field‐strength and translational spontaneous injections. Under the optimized conditions, picoliter‐scale sample plugs (corresponding to ca. 20‐μm plug length) were obtained, which ensure the high‐speed and high‐efficiency separation for DNA fragments with a short effective separation length. Other separation conditions including the sieving matrix concentration, separation field strength and effective separation length were also optimized. The present system was applied in the separation of ΦX174‐Hae III digest DNA marker. With an effective separation length of 2.5 cm, the separation could be achieved in